Steroid hormone pharmaceutical formulations

ABSTRACT

This disclosure provides pharmaceutical compositions for delivering estradiol to a subject in need thereof, as well as methods of administering the compositions, and methods of using them.

FIELD

This disclosure provides pharmaceutical compositions comprisingsolubilized estradiol. Also provided are vaginally inserted softcapsules comprising the pharmaceutical compositions and methods ofadministering the soft capsules for the treatment of vulvovaginalatrophy and female sexual dysfunction.

BACKGROUND

Postmenopausal women frequently suffer from atrophic vaginitis or vulvarand vaginal atrophy (hereafter “vulvovaginal atrophy” or “VVA”) withsymptoms including, for example, vaginal dryness, vaginal odor, vaginalor vulvar irritation or itching, dysuria (pain, burning, or stingingwhen urinating), dyspareunia (vaginal pain associated with sexualactivity), or vaginal bleeding associated with sexual activity. Othersymptoms include soreness, urinary frequency and urgency, and urinarydiscomfort and incontinence (“estrogen-deficient urinary state(s)”). Onesymptom of vaginal atrophy is an increased vaginal pH, which creates anenvironment more susceptible to infections. The cytological examinationof the vaginal mucosal epithelium in VVA subjects can also show signs ofsevere atrophy with a reduced number of superficial and intermediatecells an increased number of parabasal cells.

These symptoms are associated with decreased estrogenization of thevulvovaginal tissue and can even occur in women treated with oralestrogen-based pharmaceutical drug products. Although VVA is most commonin postmenopausal women, it can occur at any time in a woman's life, andfrequently interferes with sexual activity and satisfaction.Postmenopausal women with female sexual dysfunction (FSD) are almostfour times more likely to have VVA than those without FSD.

Estrogen treatment has proven to be very successful in controllingmenopausal symptoms, including VVA and FSD. Several studies have shownthat the symptoms associated with vulvovaginal atrophy are oftenrelieved by systemic or topical estrogen treatment.

SUMMARY

In a first aspect, the present disclosure provides a vaginal insertcomprising a therapeutically effective amount of estradiol; and asolubilizing agent, wherein the solubilizing agent comprises at leastone C₂-C₅ fatty acid or a glycol ester, monoglyceride, diglyceride, ortriglyceride thereof.

In a first embodiment of the first aspect, the therapeutically effectiveamount of estradiol is from about 1 microgram to about 25 micrograms. Ina second embodiment of the first aspect, the therapeutically effectiveamount of estradiol is from about 1 microgram to about 10 micrograms. Ina third embodiment of the first aspect, the therapeutically effectiveamount of estradiol is about 4 micrograms. In a fourth embodiment of thefirst aspect, the therapeutically effective amount of estradiol is about10 micrograms. In a fifth embodiment of the first aspect, thetherapeutically effective amount of estradiol is about 25 micrograms.

In a sixth embodiment of the first aspect, the vaginal insert furthercomprises a second solubilizing agent selected from the group consistingof a mixture of PEG-6 stearate, ethylene glycol palmitostearate, andPEG-32 stearate; a mixture of hard fat, glyceryl ricinoleate, ceteth-20,and steareth-20; polyoxyl 40 hydrogenated castor oil USP; hard fatpolyoxyl 20 cetostearyl ether; and combinations thereof. In a seventhembodiment of the first aspect, the second solubilizing agent is amixture of PEG-6 stearate, ethylene glycol palmitostearate, and PEG-32stearate.

In an eighth embodiment of the first aspect, the viscosity of thecomposition is about 5 cP to about 3000 cP at room temperature.

In a ninth embodiment of the first aspect, estradiol is the only activepharmaceutical ingredient in the vaginal insert.

In a tenth embodiment of the first aspect, the insert further comprisesa capsule encapsulating the therapeutically effective amount ofestradiol and the solubilizing agent. In an eleventh embodiment of thefirst aspect, the capsule is a soft gelatin capsule.

In a twelfth embodiment of the first aspect, the solubilizing agentcomprises at least one C₃ fatty acid or a glycol mono- or di-esterthereof, a monoglyceride, diglyceride, or triglyceride thereof, or acombination of any of the foregoing.

In a second aspect, the present disclosure provides a vaginal insertcomprising:

-   -   a therapeutically effective amount of estradiol;    -   a first solubilizing agent comprising one or more polyethylene        glycol mono- or di-esters of a hydroxy C₁₆-C₂₆ fatty acid; and    -   a second solubilizing agent comprising a mixture of PEG-6        stearate, ethylene glycol palmitostearate, and PEG-32 stearate.

In a first embodiment of the second aspect, the therapeuticallyeffective amount of estradiol is from about 1 microgram to about 25micrograms. In a second embodiment of the second aspect, thetherapeutically effective amount of estradiol is from about 1 microgramto about 10 micrograms. In a third embodiment of the second aspect, thetherapeutically effective amount of estradiol is about 4 micrograms. Ina fourth embodiment of the second aspect, the therapeutically effectiveamount of estradiol is about 10 micrograms. In a fifth embodiment of thesecond aspect, the therapeutically effective amount of estradiol isabout 25 micrograms.

In a sixth embodiment of the second aspect, estradiol is the only activepharmaceutical ingredient in the vaginal insert.

In a seventh embodiment of the second aspect, the insert furthercomprises a capsule encapsulating the estradiol, the first solubilizingagent, and the second solubilizing agent. In an eighth embodiment of thesecond aspect, the capsule is a soft gelatin capsule.

In a third aspect, the present disclosure provides a vaginal insertcomprising:

-   -   a therapeutically effective amount of estradiol;    -   a first solubilizing agent comprising propylene glycol        monolaurate; and    -   a second solubilizing agent comprising polyoxyl 40 hydrogenated        castor oil USP.

In a first embodiment of the third aspect, the therapeutically effectiveamount of estradiol is from about 1 microgram to about 25 micrograms. Ina second embodiment of the third aspect, the therapeutically effectiveamount of estradiol is from about 1 microgram to about 10 micrograms. Ina third embodiment of the third aspect, the therapeutically effectiveamount of estradiol is about 4 micrograms. In a fourth embodiment of thethird aspect, the therapeutically effective amount of estradiol is about10 micrograms. In a fifth embodiment of the third aspect, thetherapeutically effective amount of estradiol is about 25 micrograms.

In a sixth embodiment of the third aspect, the viscosity of thecomposition is about 5 cP to about 3000 cP at room temperature.

In a seventh embodiment of the third aspect, estradiol is the onlyactive pharmaceutical ingredient in the vaginal insert.

In an eighth embodiment of the third aspect, the insert furthercomprises a capsule encapsulating the estradiol, the first solubilizingagent, and the second solubilizing agent. In a ninth embodiment of thethird aspect, the capsule is a soft gelatin capsule.

In a fourth aspect, the present disclosure provides a method of treatingan estrogen-deficient state, comprising administering to a female inneed thereof, a vaginal insert comprising a therapeutically effectiveamount of estradiol; and a solubilizing agent, wherein the solubilizingagent comprises at least one C₂-C₅ fatty acid or a glycol ester,monoglyceride, diglyceride, or triglyceride thereof. In a firstembodiment of the fourth aspect, the estrogen-deficient state isvulvovaginal atrophy. In a second embodiment of the fourth aspect, theestrogen-deficient state is selected from the group consisting ofvulvovaginal atrophy, dysuria, dyspareunia, estrogen-deficient urinarystate, and vaginal bleeding associated with sexual activity.

In a third embodiment of the fourth aspect, administering the vaginalinsert comprises administering the insert daily for two weeks, and twiceweekly thereafter.

In a fifth aspect, the present disclosure provides a method of treatingan estrogen-deficient state, comprising administering to a female inneed thereof, a vaginal insert comprising:

-   -   a therapeutically effective amount of estradiol;    -   a first solubilizing agent comprising one or more polyethylene        glycol mono- or di-esters of a hydroxy C₁₆-C₂₆ fatty acid; and    -   a second solubilizing agent comprising a mixture of PEG-6        stearate, ethylene glycol palmitostearate, and PEG-32 stearate.

In a first embodiment of the fifth aspect, the estrogen-deficient stateis vulvovaginal atrophy. In a second embodiment of the fifth aspect, theestrogen-deficient state is selected from the group consisting ofvulvovaginal atrophy, dysuria, dyspareunia, estrogen-deficient urinarystate, and vaginal bleeding associated with sexual activity.

In a third embodiment of the fifth aspect, administering the vaginalinsert comprises administering the insert daily for two weeks, and twiceweekly thereafter.

In a sixth aspect, the present disclosure provides a method of treatingan estrogen-deficient state, comprising administering to a female inneed thereof, a vaginal insert comprising:

-   -   a therapeutically effective amount of estradiol;    -   a first solubilizing agent comprising propylene glycol        monolaurate; and    -   a second solubilizing agent comprising polyoxyl 40 hydrogenated        castor oil USP.

In a first embodiment of the sixth aspect, the estrogen-deficient stateis vulvovaginal atrophy. In a second embodiment of the sixth aspect, theestrogen-deficient state is selected from the group consisting ofvulvovaginal atrophy, dysuria, dyspareunia, estrogen-deficient urinarystate, and vaginal bleeding associated with sexual activity.

In a third embodiment of the sixth aspect, administering the vaginalinsert comprises administering the insert daily for two weeks, and twiceweekly thereafter.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a flow diagram illustrating a process for preparing avaginal insert.

FIG. 2 illustrates a vaginal insert in accordance with variousembodiments disclosed herein.

FIG. 3A shows an estradiol softgel capsule held with the larger endbetween the thumb and index fingers.

FIG. 3B shows insertion of an estradiol softgel capsule in a recliningposition. The softgel is inserted into the lower third of the vaginawith the smaller end up.

FIG. 3C shows insertion of an estradiol softgel capsule in a standingposition. The softgel is inserted into the lower third of the vaginawith the smaller end up.

DETAILED DESCRIPTION Definitions

The singular forms “a,” “an,” and “the” include plural referents unlessthe context clearly dictates otherwise.

As used herein, the term “or” is defined as a logical disjunction (i.e.,and/or) and does not indicate an exclusive disjunction unless expresslyindicated as such with the terms “either,” “unless,” “alternatively,”and words of similar effect.

As used herein, the term “about” refers to ±10% of the noted value,unless otherwise specified, and unless the upper bound of the rangewould exceed 100% of the pharmaceutical composition, in which case theupper limit of the range is limited to 99.9%. Thus, and by way ofexample only, a pharmaceutical composition including about 10 weightpercent of a given compound could have from 9 to 11 weight percent ofthe compound. Similarly, a pharmaceutical composition including about 95weight percent of a given compound could have from 85.5 to 99.9 weightpercent of the compound in the pharmaceutical composition.

As used herein, the term “drug product” means at least one activepharmaceutical ingredient in combination with at least one excipient andprovided in unit dosage form.

As used herein, the term “hormone deficiency” refers to a low level ofone or more steroid hormones in a subject. Normal hormone levels willvary from subject to subject and can be determined via known methods.Low hormone levels may or may not be associated with symptoms including,for example and without limitation, vasomotor symptoms, sleepdisturbances, mood changes, and vulvovaginal atrophy.

As used herein, the term “subject” refers to any animal, includinghumans.

The term “micronized” as used herein, refers to particles having an X50particle size value below about 15 microns or having an X90 particlesize value below about 25 microns. In some embodiments, a micronizedparticle can have an X90 particle size of less than 5 microns. The term“X50” means that one-half of the particles in a sample are smaller indiameter than a given number. For example, a micronized particle havingan X50 of 5 microns means that, for a given sample of the micronizedparticle, one-half of the particles have a diameter of less than 5microns. Similarly, the term “X90” means that ninety percent (90%) ofthe particles in a sample are smaller in diameter than a given number.

As used herein, the term “predominantly” means at least 50 percent. Byway of example only, a compound comprising a linear predominantly C10alkylene group, comprises at least 50 percent, at least 60 percent, atleast 70 percent, at least 80 percent, at least 85 percent, at least 90percent, at least 91 percent, at least 92 percent, at least 93 percent,at least 94 percent, at least 95 percent, at least 96 percent, at least97 percent, at least 98 percent, or at least 99 percent of the linearC10 alkylene group, with the remainder being an alkylene group eithergreater than or less than C10. In certain embodiments, predominantlymeans at least 85 percent. “Predominantly” can be used in a variety ofunit measurement systems, including mol %, w/w, or aggregate number offatty acid esters, for example.

The term “glyceride” is an ester of glycerol (1,2,3-propanetriol) withacyl radicals of fatty acids and is known as an acylglycerol. If onlyone position of the glycerol molecule is esterified with a fatty acid, a“monoglyceride” or “monoacylglycerol” is produced; if two positions areesterified, a “diglyceride” or “diacylglycerol” is produced; and if allthree positions of the glycerol are esterified with fatty acids, a“triglyceride” or “triacylglycerol” is produced. A glyceride is “simple”if all esterified positions contain the same fatty acid; whereas aglyceride is “mixed” if the esterified positions contained differentfatty acids. The carbons of the glycerol backbone are designated sn-1,sn-2 and sn-3, with sn-2 being in the middle carbon and sn-1 and sn-3being the end carbons of the glycerol backbone.

The term “solubilizing agent” refers to an agent or combination ofagents that solubilize an active pharmaceutical ingredient (e.g.,estradiol). Solubilizing agents include agents that solubilize ordissolve an active pharmaceutical ingredient to a desirable extent.Solubilizing agents suitable for use in the formulations disclosedherein are pharmaceutical grade solubilizing agents. It will beunderstood by those of skill in the art that other excipients orcomponents can be added to or mixed with the solubilizing agent toenhance the properties or performance of the solubilizing agent orresulting formulation. Examples of such excipients include, but are notlimited to, surfactants, emulsifiers, thickeners, colorants, flavoringagents, etc.

As used herein, the term “prevent” refers to the prophylactic treatmentof a subject who is at risk of developing a condition (e.g., steroidhormone deficiency) resulting in a decrease in the probability that thesubject will develop the condition.

The terms “treat,” “treating,” “treatment” and the like refer to anyindicia of success in the treatment or amelioration of an injury,disease, or condition, including any objective or subjective parametersuch as abatement; remission; diminishing of symptoms or making theinjury, disease, or condition more tolerable to the subject; slowing inthe rate of degeneration or decline; or improving a subject's physicalor mental well-being. The treatment or amelioration of symptoms can bebased on objective or subjective parameters, including the results of aphysical examination, neuropsychiatric examinations, or psychiatricevaluation.

The terms “atrophic vaginitis,” “vulvovaginal atrophy,” “vaginalatrophy,” and “VVA” are used herein interchangeably. The cellularmorphology of VVA is well known in the medical field.

As used herein, “sexual dysfunction” refers to a condition having one ormore symptoms or difficulties involving one or more aspects of sexualactivity. The dysfunction can prevent an individual from enjoying sexualactivity. Nonlimiting examples of symptoms of sexual dysfunctioninclude, but are not limited to, dyspareunia, painful contractions(spasms) of the vaginal muscles, and problems with sexual desire,arousal, or orgasm. Sexual dysfunction can be lifelong (no effectiveperformance) or acquired (after a period of normal function); and can begeneralized or limited to certain situations or certain partners.

As used herein, “dyspareunia” refers to persistent or recurrent genitalpain that occurs just before, during, or after sexual activity.

As used herein, “lubrication” refers to wetness in and around the vaginabefore, during, or after sexual activity. Increasing lubrication caninclude increasing the frequency of lubrication; decreasing thedifficulty of becoming lubricated; and/or decreasing the difficulty inmaintaining lubrication.

As used herein, “sexual desire” refers to the frequency of wanting toengage in sexual activity and/or the frequency of engaging in sexualactivity as perceived by the individual. Sexual desire can be expressed,for example, in one or more cognitive activities, including thefrequency of sexual thoughts, the extent of enjoyment of movies, books,music, etc. having sexual content and/or the extent of enjoyment orpleasure of thinking and fantasizing about sex as perceived by theindividual.

As used herein, “sexual arousal” refers to the frequency of becomingsexually aroused, how readily sexual arousal occurs and/or if arousal ismaintained, as perceived by the individual. Psychologically, arousal caninclude factors such as increased desire for sexual activity andexcitement related to sexual activity. Physiologically, arousal caninclude increased blood flow to the genitals, causing clitoralengorgement, as well as vaginal lubrication.

As used herein, “orgasm” refers to the highest point of sexualexcitement characterized by a subjective experience of intense pleasuremarked normally by vaginal contractions in females. Increasing orgasmcan include increasing the frequency, duration, and/or intensity oforgasms in a subject. Increasing orgasm can also include decreasing thedifficulty of reaching orgasm.

As used herein, “satisfaction” refers to one or more positive emotions(e.g., contentment, fulfillment, gratification, and the like) related toa sexual activity or sexual relationship. Satisfaction can include, forexample, satisfaction with occurrence of sexual arousal or orgasm,satisfaction with the amount of closeness with a partner, andsatisfaction with overall sex life.

The phrase “therapeutically effective amount” refers to an amount of apharmaceutical composition or of a given steroid hormone suitable totreat a particular symptom, disorder, or disease.

As used herein, the phrase “substantially” means at least about 90%, incertain embodiments, at least about 95%, and in still furtherembodiments, at least about 98%. For example, an object that is“substantially pure” or an object that is “substantially free” ofanother object, refers to a compound or composition that is at leastabout 90% pure by weight, at least about 95% pure by weight, or at leastabout 98% pure by weight and contains less than about 10% by weight,less than about 5% by weight or less than about 2% by weight ofcontaminants.

As used herein, the phrase “steroid hormone” refers to estradiol.

The term “bio-identical,” “body-identical,” or “natural” used inconjunction with the hormones disclosed herein, means hormones thatmatch the chemical structure and effect of those that occur naturally orendogenously in the human body. An exemplary natural estrogen is17β-estradiol.

The term “estradiol” refers to (17β)-estra-1,3,5(10)-triene-3,17-diol.Estradiol is also interchangeably called 17β-estradiol, oestradiol, orE2, and is found endogenously in the human body. As used herein,estradiol refers to the bio-identical or body-identical form ofestradiol found in the human body having the structure:

Estradiol is supplied in an anhydrous or hemi-hydrate form. For thepurposes of this disclosure, the anhydrous form or the hemihydrate formcan be substituted for the other by accounting for the water or lack ofwater according to well-known and understood techniques.

The term “solubilized estradiol” means that the estradiol or a portionthereof is solubilized or dissolved in the solubilizing agent(s) or theformulations disclosed herein. Solubilized estradiol can includeestradiol that is about 80% solubilized, about 85% solubilized, about90% solubilized, about 95% solubilized, about 96% solubilized, about 97%solubilized, about 98% solubilized, about 99% solubilized, or about 100%solubilized. In some embodiments, the estradiol is “fully solubilized”with all or substantially all of the estradiol being solubilized ordissolved in the solubilizing agent. Fully solubilized estradiol caninclude estradiol that is about 97% solubilized, about 98% solubilized,about 99% solubilized or 100% solubilized. Solubility can be expressedas a mass fraction (% w/w, which is also referred to as weight percent(wt %)).

The solubility of a given steroid hormone can be measured using standardtechniques by weighing a piece of filter paper, placing the weighedfilter paper in a Buchner funnel (porcelain or glass with a glass frit),and drawing a known quantity of pharmaceutical composition through thefilter paper using vacuum (such as with a side-arm flask fitted with aneoprene collar). After drying for an appropriate period of time (eitherat room temperature or at elevated temperature), the filter paper isreweighed. The amount of steroid hormone on the filter paper iscalculated and the amount of soluble and insoluble steroid hormone iscalculated.

The term “excipients,” as used herein, refers to non-API ingredientssuch as solubilizing agents, anti-oxidants, oils, lubricants, and otheragents used in formulating pharmaceutical products.

The term “active pharmaceutical ingredient” (“API”) as used herein,means the active compound(s) used in formulating a drug product.

Pharmaceutical Compositions

Provided herein are pharmaceutical compositions comprising estradioldesigned to be absorbed vaginally and have a local therapeutic effect(e.g., in vaginal and vulvar tissues). Generally, the pharmaceuticalcompositions disclosed herein are useful for treating VVA, dyspareunia,and other disorders caused by decreased estrogen presence.

Functionality

In certain embodiments, the pharmaceutical compositions disclosed hereincan be alcohol-free or substantially alcohol-free. In certainembodiments, the pharmaceutical compositions disclosed herein can beencapsulated in soft gelatin capsules.

Estradiol

In certain embodiments, the pharmaceutical compositions disclosed hereinare for vaginal insertion through a single dose or through multipledoses. In certain embodiments, the estradiol in the pharmaceuticalcompositions can be 100% solubilized. In certain embodiments, theestradiol in the pharmaceutical compositions can be less than 100%solubilized. In embodiments where the estradiol is less than 100%solubilized, the remaining estradiol can be present in a micronized(crystalline) form that is absorbable by the body and retains biologicalfunctionality, either in its micronized form or in another form whichthe micronized form is converted to after administration.

In certain embodiments, all or some of the estradiol can be solubilizedin one or more solubilizing agents during manufacturing process. Incertain embodiments, all or some of the estradiol can be solubilizedfollowing administration (e.g., insoluble estradiol in the formulationcan be solubilized in a body fluid after administration). To the extentthe estradiol is not fully solubilized at the time ofadministration/insertion, the estradiol will be substantiallysolubilized at a body temperature (average of 37° C.) and, generally, atthe pH of the vagina (ranges from 3.8 to 4.5 in healthy subjects; or 4.6to 6.5 in VVA subjects).

In certain embodiments, estradiol can be soluble in the solubilizingagent(s) at room temperature, although it can be desirable to warmcertain solubilizing agents during manufacture to improve viscosity. Incertain embodiments, the solubilizing agent can be a liquid orsemi-solid. In certain embodiments, the solubilizing agent can be aliquid at between room temperature and about 50° C., at or below 50° C.,at or below 40° C., or at or below 30° C.

In certain embodiments, the viscosity of the pharmaceutical compositionsdescribed this disclosure can range from about 5 centipose (“cP”) toabout 3000 cP at 25° C. A person of ordinary skill in the art canreadily understand and select from suitable solubilizing agents toarrive at the desired viscosity. Viscosity can be measured by varioustypes of viscosity meters and rheometers. These instruments measure thefriction between the fluid and an adjacent layer. For example, aBrookfield Viscometer (DVEELVTJ0) can be used to measure the viscosityof these compositions by using an S31 spindle at 5-100 rpm and at roomtemperature.

In certain embodiments, the viscosity of the pharmaceutical compositionsdescribed in this disclosure will exceed 3000 cP, e.g., for semi-solidsor solids. A person of ordinary skill in the art can readily measure theviscosity of such semi-solids or solids by using other known methods,e.g., increasing temperature or using different spindles.

In certain embodiments, the solubility of estradiol in the solubilizingagent(s) can be about 0.01 wt %, 0.02 wt %, 0.05 wt %, 0.06 wt %, 0.08wt %, 0.1 wt %, 0.2 wt %, 0.3 wt %, 0.4 wt %, 0.5 wt %, 0.6 wt %, 0.7 wt%, 0.8 wt %, 0.9 wt %, 1.0 wt %, or higher. In some embodiments, thesolubility of estradiol in the solubilizing agent(s) can be from about0.001 wt % to about 5 wt %, from about 0.001 wt % to about 2 wt %, fromabout 0.001 wt % to about 1 wt %, from about 0.001 wt % to about 0.5 wt%, from about 0.001 wt % to about 0.1 wt %, from about 0.001 wt % toabout 0.05 wt %, from about 0.01 wt % to about 5 wt %, from about 0.01wt % to about 2 wt %, from about 0.01 wt % to about 1 wt %, from about0.01 wt % to about 0.5 wt %, from about 0.01 wt % to about 0.1 wt %,from about 0.1 wt % to about 5 wt %, from about 0.1 wt % to about 2 wt%, from about 0.1 wt % to about 1 wt %, from about 0.1 wt % to about 0.5wt %, from about 0.5 wt % to about 5 wt %, from about 0.5 wt % to about2 wt %, from about 0.5 wt % to about 1 wt %, from about 1 wt % to about5 wt %, from about 1 wt % to about 2 wt %, or from about 2 wt % to about5 wt %.

In certain embodiments, the estradiol can be added to the pharmaceuticalcompositions disclosed herein as estradiol, estradiol hemihydrate, orany other estradiol form(s) suitable for use in pharmaceuticalcompositions or formulations.

The pharmaceutical compositions described herein can contain estradiolin varying amounts. Estradiol (or estradiol hemihydrate, for example, tothe extent the water content of the estradiol hemihydrate is accountedfor) can be present in the formulation in a dosage amount ranging fromabout 1 microgram (m) to about 50 μg. In other embodiments, thepharmaceutical composition can contain about: 1 μg, 2 μg, 3 μg, 4 μg, 5μg, 6 μg, 7 μg, 8 μg, 9 μg, 10 μg, 11 μg, 12 μg, 13 μg, 14 μg, 15 μg, 16μg, 17 μg, 18 μg, 19 μg, 20 μg, 21 μg, 22 μg, 23 μg, 24 μg, 25 μg, 26μg, 27 μg, 28 μg, 29 μg, 30 μg, 31 μg, 32 μg, 33 μg, 34 μg, 35 μg, 36μg, 37 μg, 38 μg, 39 μg, 40 μg, 41 μg, 42 μg, 43 μg, 44 μg, 45 μg, 46μg, 47 μg, 48 μg, 49 μg, or 50 μg estradiol (or an equivalent amount of,for example, estradiol hemihydrate). In certain embodiments, thepharmaceutical composition can contain about 2.5 μg, 4 μg, 6.25 μg, 7.5μg, 12.5 μg, or 18.75 μg of estradiol. In certain embodiments, thepharmaceutical composition can contain from about 1 μg to about 10 fromabout 3 μg to about 7 μg, from about 7.5 μg to 12.5 μg, from about 10 μgto about 25 μg, from about 1 μg, about 2.5 μg, from about 23.5 μg toabout 27.5 μg, from about 7.5 μg to about 22.5 μg, or from about 10 μgto about 25 μg of estradiol. In one embodiment, the pharmaceuticalcomposition comprises about 4 μg of estradiol. In one embodiment, thepharmaceutical composition comprises about 10 μg of estradiol. Inanother embodiment, the pharmaceutical composition comprises about 25 μgof estradiol.

In some embodiments, estradiol is the only active hormone in thevaginally inserted pharmaceutical composition.

Short Chain (C₂-C₅) Fatty Acids as Solubilizing Agents

In certain embodiments, the solubilizing agent can comprise at least oneshort chain fatty acid (C₂-C₅). In some embodiments, the short chainfatty acid can be acetic acid (C₂), propionic acid (C₃), butyric acid(C₄), isobutyric acid (C₄), valeric acid (C₅), isovaleric acid (C₅), ormixtures thereof.

In embodiments where the solubilizing agent comprises one or more C₃-C₅short chain fatty acids, the one or more C₃-C₅ fatty acids can besaturated. In certain embodiments, the one or more C₃-C₅ short chainfatty acids can be predominantly saturated, i.e., greater than about 60%or greater than about 75% saturated.

In certain embodiments, the solubilizing agent can comprise one or morefree short chain fatty acids, one or more short chain fatty acid (C₂-C₅)esters of glycerin, propylene glycol, ethylene glycol, or combinationsthereof. Exemplary short chain fatty acid (C₂-C₅) esters of glycerininclude, but are not limited to, monoacetylglyceride (MAG),diacetylglyceride (DAG), triacetin (triacetylglyceride, TAG), propionictriglyceride, monobutylglyceride, dibutylglyceride, butyric triglyceride(tributyrin), isobutyric triglyceride, isobutyric diglyceride,isobutyric monoglyceride, valeric triglyceride, valeric diglyceride,valeric monoglyceride, isovaleric monoglyceride, isovaleric diglycerideand isovaleric triglyceride. In some embodiments, the solubilizing agentcan comprise triacetin.

In certain embodiments, the pharmaceutical composition can comprise afirst solubilizing agent comprising at least one C₂-C₅ fatty acid or aglycol ester, monoglyceride, diglyceride, or triglyceride thereof and asecond solubilizing agent. The second solubilizing agent can be anyagent that solubilizes or dissolves the steroid hormone to the desirableextent.

In some embodiments, the second solubilizing agent can be TEFOSE 63(mixture of PEG-6 stearate and ethylene glycol palmitostearate andPEG-32 stearate, available from GATTEFOSSÉ SAS, Saint-Priest, France),TEFOSE 1500 (mixture of PEG-6 stearate and PEG-32 stearate, availablefrom GATTEFOSSÉ SAS, Saint-Priest, France), TEFOSE 2000 (mixture ofPEG-6 stearate, ceteth-20, and stearath-20, available from GATTEFOSSÉSAS, Saint-Priest, France), or GELOT 64 (mixture of PEG-75 stearate andglyceryl stearate, available from GATTEFOSSÉ SAS, Saint-Priest, France).

In certain embodiments, the second solubilizing agent can comprise oneor more C₆-C₁₈ fatty acid PEG mono- or di-esters, one or more C₆-C₁₈fatty acid mono-, di-, or tri-esters of glycerol, or combinationsthereof. In one embodiment, the second solubilizing agent can beGELUCIRE 44/14 (lauroyl macrogol-32 glycerides (EP); lauroyl polyoxyl-32glycerides (USP-NF)). Gelucire 44/14 is well known in the art andcomprises a small fraction of mono-, di-, and triglycerides and a mainfraction comprising PEG-32 (MW 1500) mono- and diesters of lauric acid(C12).

In certain embodiments, the second solubilizing agent can comprise acastor oil or hydrogenated castor oil ethoxylate. In particularembodiments, the castor oil or hydrogenated castor oil ethoxylate can beKOLLIPHOR EL (polyethoxylated castor oil, available from BASF),KOLLIPHOR RH40 (polyoxyl 40 hydrogenated castor oil USP, available fromBASF), MYRJ 52 (polyoxyl 40 stearate, available from Spectrum Chemical),ETOCAS 40 (polyoxyethylene (40) castor oil, available from Croda),LABRASOL (caprylocaproyl polyoxyl-8-glycerides NF and caprylocaproylmacrogol-8-glycerides EP, available from GATTEFOSSÉ SAS, Saint-Priest,France), or CRODURET 60 (PEG 60 hydrogenated castor oil, available fromCroda).

In certain embodiments, the second solubilizer can comprise OVUCIRE 3460(a mixture of hard fat, glyceryl ricinoleate, ceteth-20, andsteareth-20, available from GATTEFOSSÉ, Saint-Priest, France) or OVUCIREWL 3264 (hard fat polyoxyl 20 cetostearyl ether, available fromGATTEFOSSÉ, Saint-Priest, France).

In some embodiments, the weight:weight ratio of the first solubilizingagent to the second solubilizing agent can range from about 4:1 to about10:1. In particular embodiments, the ratio of first solubilizing agentto second solubilizing agent is about 9:1.

In some embodiments, the pharmaceutical composition can comprise a firstsolubilizing agent comprising at least one C₂-C₅ fatty acid or a glycolester, monoglyceride, diglyceride, or triglyceride thereof, and a secondsolubilizing agent comprising a mixture polyoxylglycerides and glycols.In particular embodiments, the pharmaceutical composition can comprise a9:1 mixture of triacetin and TEFOSE 63 (mixture of PEG-6 stearate andethylene glycol palmitostearate and PEG-32 stearate, available fromGATTEFOSSÉ SAS, Saint-Priest, France).

Hydroxy Fatty Acids as Solubilizing Agents

In certain embodiments, the solubilizing agent can comprise at least onehydroxy fatty acid, such as at least one hydroxy C₁₆-C₂₆ fatty acid, atleast one hydroxy C₁₆ to C₂₄ fatty acid, at least one hydroxy C₁₆ to C₂₂fatty acid, at least one hydroxy C₁₆ to C₂₀, at least one hydroxy C₁₆ toC₁₈ fatty acid, at least one hydroxy C₁₈ to C₂₆ fatty acid, at least onehydroxy C₁₈ to C₂₄ fatty acid, at least one hydroxy C₁₈ to C₂₂ fattyacid, at least one hydroxy C₁₈ to C₂₀ fatty acid, or a combinationthereof.

In some embodiments, the various suitable hydroxy fatty acids notedabove can be saturated. In some embodiments, these hydroxy fatty acidscan be predominantly saturated, i.e., greater than about 60% or greaterthan about 75% saturated.

In certain embodiments, the hydoxy fatty acid can be a C₁₆-C₂₆ hydoxyfatty acid and can be α-hydroxymyristic acid, β-hydroxymyristic acid,3-hydroxy-13-methyltetradecanoic acid, α-hydroxypalmitic acid,β-hydroxypalmitic acid, 3-hydroxy-15-methylhexadecanoic acid,β-hydroxystearic acid, 12-hydroxystearic acid, and 17-hydroxystearicacid, or a mixture of any of the foregoing.

In certain embodiments, the solubilizing agent can comprise one or moreglycol mono- or di-esters of a hydroxy C₁₆-C₂₆ fatty acid. Suitableglycols can include ethylene glycol, propylene glycol, and polyethyleneglycol. In some embodiments, the solubilizing agent can comprise one ormore polyethylene glycol mono- or di-esters of a hydroxy C₁₆-C₂₆ fattyacid. In some embodiments, the solubilizing agent can comprise one ormore polyethylene glycol mono- or di-esters of a C₁₆-C₂₆ fatty acid. Insome embodiments, the solubilizing agent comprises a mixture of one ormore polyethylene glycol mono- and di-esters of a hydroxy C₁₆-C₂₆ fattyacid and one or more polyethylene glycol di-esters of a C₁₆-C₂₆ fattyacid. In some embodiments, the solubilizing agent comprises polyethyleneglycol mono- and di-esters of 12-hydroxystearic acid and freepolyethylene glycol (sold as KOLLIPHOR HS 15, available from BASF).

In certain embodiments, the pharmaceutical composition can comprise afirst solubilizing agent comprising one or more polyethylene glycolmono-esters of a C₆-C₂₆ hydroxyfatty acid, one or more polyethyleneglycol di-esters of a C₆-C₂₆ hydroxyfatty acid, or a combinationthereof; and a second solubilizing agent.

In some embodiments, the second solubilizing agent can be TEFOSE 63(mixture of PEG-6 stearate and ethylene glycol palmitostearate andPEG-32 stearate, available from GATTEFOSSÉ SAS, Saint-Priest, France),TEFOSE 1500 (mixture of PEG-6 stearate and PEG-32 stearate, availablefrom GATTEFOSSÉ SAS, Saint-Priest, France), TEFOSE 2000 (mixture ofPEG-6 stearate, ceteth-20, and stearath-20, available from GATTEFOSSÉSAS, Saint-Priest, France), or GELOT 64 (mixture of PEG-75 stearate andglyceryl stearate, available from GATTEFOSSÉ SAS, Saint-Priest, France).

In certain embodiments, the second solubilizing agent can comprise oneor more C₆-C₁₈ fatty acid PEG mono- or di-esters, one or more C₆-C₁₈fatty acid mono-, di-, or tri-esters of glycerol, and combinationsthereof. In one embodiment, the second solubilizing agent can beGELUCIRE 44/14.

In certain embodiments, the second solubilizing agent can comprise acastor oil or hydrogenated castor oil ethoxylate. In particularembodiments, the castor oil or hydrogenated castor oil ethoxylate can beKOLLIPHOR EL (polyethoxylated castor oil, available from BASF),KOLLIPHOR RH40 (polyoxyl 40 hydrogenated castor oil USP, available fromBASF), MYRJ 52 (polyoxyl 40 stearate, available from Spectrum Chemical),ETOCAS 40 (polyoxyethylene (40) castor oil, available from Croda),LABRASOL (caprylocaproyl polyoxyl-8-glycerides NF and caprylocaproylmacrogol-8-glycerides EP, available from GATTEFOSSÉ SAS, Saint-Priest,France), or CRODURET 60 (PEG 60 hydrogenated castor oil, available fromCroda).

In certain embodiments, the second solubilizer can comprise OVUCIRE 3460(a mixture of hard fat, glyceryl ricinoleate, ceteth-20, andsteareth-20, available from GATTEFOSSÉ, Saint-Priest, France) or OVUCIREWL 3264 (hard fat polyoxyl 20 cetostearyl ether, available fromGATTEFOSSÉ, Saint-Priest, France).

In some embodiments, the weight:weight ratio of the first solubilizingagent to the second solubilizing agent can range from 4:1 to about 10:1.In particular embodiments, the ratio of first solubilizing agent tosecond solubilizing agent can be about 9:1.

In some embodiments, the pharmaceutical composition can comprise a firstsolubilizing agent comprising one or more PEG esters of a C₆-C₂₆ hydroxyfatty acid, one or more PEG diesters of a C₆-C₂₆ hydroxy fatty acid, ora combination thereof; and a second solubilizing agent comprising amixture polyoxylglycerides and glycols. In particular embodiments, thepharmaceutical composition can comprise a 9:1 (w:w) mixture of KOLLIPHORHS 15 (polyethylene glycol mono- and di-esters of 12-hydroxystearic acidand free polyethylene glycol, available from BASF) and TEFOSE 63(mixture of PEG-6 stearate and ethylene glycol palmitostearate andPEG-32 stearate, available from GATTEFOSSÉ SAS, Saint-Priest, France).

Propylene Glycol Monolaurate as a Solubilizing Agent

In certain embodiments, the solubilizing agent can comprise propyleneglycol monolaurate (sold as LAUROGLYCOL 90, available from GATTEFOSSÉSAS, Saint-Priest, France).

In some embodiments, the pharmaceutical composition can comprisepropylene glycol monolaurate (sold as LAUROGLYCOL 90, available fromGATTEFOSSÉ SAS, Saint-Priest, France) and a second solubilizing agent.

In some embodiments, the second solubilizing agent can be TEFOSE 63(mixture of PEG-6 stearate and ethylene glycol palmitostearate andPEG-32 stearate, available from GATTEFOSSÉ SAS, Saint-Priest, France),TEFOSE 1500 (mixture of PEG-6 stearate and PEG-32 stearate, availablefrom GATTEFOSSÉ SAS, Saint-Priest, France), TEFOSE 2000 (mixture ofPEG-6 stearate, ceteth-20, and stearath-20, available from GATTEFOSSÉSAS, Saint-Priest, France), or GELOT 64 (mixture of PEG-75 stearate andglyceryl stearate, available from GATTEFOSSÉ SAS, Saint-Priest, France).

In certain embodiments, the second solubilizing agent can comprise oneor more C₆-C₁₈ fatty acid PEG mono- or di-esters, one or more C₆-C₁₈fatty acid mono-, di-, or tri-esters of glycerol, and combinationsthereof. In one embodiment, the second solubilizing agent can beGELUCIRE 44/14.

In certain embodiments, the second solubilizing agent can comprise acastor oil or hydrogenated castor oil ethoxylate. In particularembodiments, the castor oil or hydrogenated castor oil ethoxylate can beKOLLIPHOR EL (polyethoxylated castor oil, available from BASF),KOLLIPHOR RH40 (polyoxyl 40 hydrogenated castor oil USP, available fromBASF), MYRJ 52 (polyoxyl 40 stearate, available from Spectrum Chemical),ETOCAS 40 (polyoxyethylene (40) castor oil, available from Croda),LABRASOL (caprylocaproyl polyoxyl-8-glycerides NF and caprylocaproylmacrogol-8-glycerides EP, available from GATTEFOSSÉ SAS, Saint-Priest,France), or CRODURET 60 (PEG 60 hydrogenated castor oil, available fromCroda).

In certain embodiments, the second solubilizer can comprise OVUCIRE 3460(a mixture of hard fat, glyceryl ricinoleate, ceteth-20, andsteareth-20, available from GATTEFOSSÉ, Saint-Priest, France) or OVUCIREWL 3264 (hard fat polyoxyl 20 cetostearyl ether, available fromGATTEFOSSÉ, Saint-Priest, France).

In some embodiments, the weight:weight ratio of the first solubilizingagent to the second solubilizing agent can be from 4:1 to about 10:1. Inparticular embodiments, the ratio of first solubilizing agent to secondsolubilizing agent can be about 9:1 (w:w).

In some embodiments, the pharmaceutical composition can comprisepropylene glycol monolaurate (sold as LAUROGLYCOL 90, available fromGATTEFOSSÉ SAS, Saint-Priest, France) and a second solubilizing agentcomprising polyoxyl 40 hydrogenated castor oil (sold as KOLLIPHOR RH40,available from BASF). In particular embodiments, the pharmaceuticalcomposition can comprise a 9:1 mixture of LAUROGLYCOL 90 and KOLLIPHORRH40

Methods of Treating Hormone Deficiencies

In addition to the compositions described above, this disclosure furtherprovides methods for treating one or more conditions associated withhormone deficiency in a subject. The methods comprise vaginallyadministering to a subject in need thereof an effective amount of thepharmaceutical composition described herein.

In some embodiments, the condition being treated can be an estrogendeficiency in the vulvovaginal area, including conditions such as VVA.

In certain embodiments, the pharmaceutical composition can beadministered once daily within in any of the above noted amounts untilthe disease or condition is treated.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein administering the vaginal insert does not require use of anapplicator.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein administering the vaginal insert is by digital insertion.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein administering the vaginal insert is by insertion using anapplicator.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein the vaginal insert is inserted about two inches into the vagina.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein administering the vaginal insert results in no or minimalsubject irritation.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein subject irritation is selected from the group consisting ofpain, itching, soreness, excessive discharge, swelling, or combinationsthereof.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein subject irritation associated with administering the vaginalinsert containing the API does not differ significantly from subjectirritation associated with administering a placebo insert.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein subject irritation associated with administering the vaginalinsert is reduced compared to subject irritation associated withadministering other hormone replacement vaginal inserts.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein after two weeks of administration, the frequency ofadministering the vaginal insert is less frequent than that required byother hormone replacement vaginal inserts.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein time to ambulation for a subject is less than 20 minutes afteradministering the vaginal insert.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein time to ambulation for a subject is less than 10 minutes afteradministering the vaginal insert.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein time to ambulation for a subject is less than 5 minutes afteradministering the vaginal insert.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein time to ambulation for a subject after administering the vaginalinsert is reduced compared to time to ambulation associated withadministering other hormone replacement vaginal inserts.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein the vaginal insert can be administered when substantially novaginal fluid is present in the vagina.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein the vaginal insert further comprises a capsule.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein the vaginal insert further comprises a capsule, wherein thecapsule is substantially dissolved in the vagina. In some embodiments,at least about 90% of the capsule is dissolved in the vagina.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein the vaginal insert further comprises a capsule, wherein there issubstantially no discharge of contents of the vaginal insert observedwithin 30 minutes after administering the vaginal insert.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein the vaginal insert further comprises a capsule, wherein there issubstantially no discharge of contents of the vaginal insert observedwithin 2 hours after administering the vaginal insert.

In some embodiments, the method of treating the symptoms of VVAcomprises administering a vaginal insert comprising the pharmaceuticalcomposition described herein to the vagina of a subject in need thereof,wherein the vaginal insert further comprises a capsule, wherein theamount of discharge of contents of the vaginal insert is less than theamount of discharge observed with other hormone replacement vaginalinserts, as observed by visual examination.

Also provided is a method for enhancing subject compliance in apopulation of subjects having symptoms of VVA, comprising administeringa vaginal insert comprising the pharmaceutical composition describedherein, wherein the pharmaceutical composition comprises 1 microgram to25 micrograms of estradiol, to the vagina of a subject in need thereof,wherein the subject administers the vaginal insert at least daily fortwo weeks.

In some embodiments, the present disclosure provides a method forenhancing subject compliance in a population of subjects having symptomsof VVA, the method comprising administering a vaginal insert comprisingthe pharmaceutical composition described herein to the vagina of asubject in need thereof, wherein the subject administers the vaginalinsert daily for two weeks, and at least twice weekly thereafter.

In some embodiments, the present disclosure provides a method forenhancing subject compliance in a population of subjects having symptomsof VVA, the method comprising administering a vaginal insert comprisingthe pharmaceutical composition described herein to the vagina of asubject in need thereof, wherein compliance of subjects in thepopulation administering the vaginal insert is enhanced compared tocompliance of subjects in a population administering other hormonereplacement vaginal inserts.

In certain embodiments, the present disclosure provides a method forestrogenizing the vulva, which consists of the labia majora, the labiaminora (labia majora and minora are collectively referred to as “thelabia”), clitoris, vestibule of the vagina, bulb of the vestibule, andthe glands of Bartholin (all of which can come into contact with theliquid that partially flows from the vagina). In some embodiments, anestradiol containing pharmaceutical composition can be digitallyinserted about two inches into the vagina or inserted into the third ofthe vagina closest to the vaginal opening as shown in FIGS. 3A, 3B, and3C. Without wishing to be bound to a particular theory, it is believedthat the estradiol containing pharmaceutical composition dissolves,ruptures, or otherwise releases the pharmaceutical composition into thevagina, whereby the lower third of the vagina and labia are bothreestrogenized. In some embodiments, the pharmaceutical composition is aliquid that partially flows to the labia and directly reestrogenizes thelabia.

In certain embodiments, the present disclosure provides a method foravoiding transport of the estradiol to the uterus comprisingadministering an estradiol containing composition into the lower thirdof the vagina closest to the vaginal opening as shown in FIGS. 3A, 3B,and 3C. Without wishing to be bound by a particular theory, it isbelieved that the estradiol containing composition releases theestradiol in the lower third of the vagina, which substantiallyeliminates transport of the estradiol to the uterus, where unopposedestradiol can cause endometrial hyperplasia, which could potentiallylead to endometrial cancer.

In certain embodiments, the present disclosure provides a method forestrogenizing the vulva. In some embodiments, the pharmaceuticalcomposition is digitally inserted about two inches into the vagina orinserted into the third of the vagina closest to the vaginal opening asshown in FIGS. 3A, 3B, and 3C. Without wishing to be bound to aparticular theory, it is believed that the gelatin capsule containingthe pharmaceutical composition dissolves, ruptures, or otherwisereleases the pharmaceutical composition into the vagina, whereby thelower third of the vagina and vulva are both reestrogenized. In someembodiments, the pharmaceutical composition can be a liquid thatpartially flows to the vulval tissue and directly reestrogenizes thevulva.

In certain embodiments, the present disclosure provides a method fortreating VVA, including dyspareunia, vaginal dryness, andestrogen-deficient urinary states (including urinary tract infections),wherein a delivery vehicle comprising the pharmaceutical composition fortreating VVA is digitally inserted approximately two inches into thevagina or into the third of the vagina closest to the opening of thevagina. This method can result in at least one of: improved compliancecompared to other products for the treatment of VVA; improved userexperience compared to other products for the treatment of VVA; andstatistically significantly improved symptoms of VVA, compared toplacebo or baseline within at least one, two, four, six, eight, ten,twelve, or more weeks after initiating administration. In certainembodiments, a method for the treatment of VVA, including dyspareunia,vaginal dryness, and estrogen-deficient urinary states (includingurinary tract infections), is provided wherein a delivery vehiclecontaining a composition for the treatment of VVA in a tear drop shapedcapsule as disclosed herein is inserted approximately two inches intothe vagina or into the third of the vagina closest to the opening of thevagina. This method can result in at least one of: improved compliancecompared to other products for the treatment of VVA; improved userexperience compared to other products for the treatment of VVA; andstatistically significantly improved symptoms of VVA, compared toplacebo or baseline within at least one, two, four, six, eight, ten,twelve, or more weeks after initiation of administration.

Methods for Preparing the Pharmaceutical Compositions

In certain embodiments, the pharmaceutical composition can be preparedby blending estradiol with at least one pharmaceutically acceptablesolubilizing agent. In certain embodiments, other excipients including,for example and without limitation, nontoxic pharmaceutically acceptablesolvents, co-solvents, surfactants, lubricants, antioxidants, and/orother excipients suitable for vaginal delivery or absorption, and thelike can be added to the pharmaceutical composition. In certainembodiments, the pharmaceutical composition can include sufficientsolubilizing agent to fully solubilize the estradiol. It is expresslyunderstood, however, the other volumes of solubilizing agent can be useddepending on the level of estradiol solubilization desired. Persons ofordinary skill in the art will know and understand how to determine thevolume of solubilizing agent and other excipients depending on thedesired percent of estradiol to be solubilized in the pharmaceuticalcomposition. In some embodiments, the solubilizing agent can be heated(for example, from about 40° C. to about 65° C.), although such heatingis not necessary to dissolve the estradiol.

Delivery Vehicle

Generally, the pharmaceutical compositions described herein aredelivered intravaginally inside of a delivery vehicle, for example acapsule. In certain embodiments, the capsules are soft capsules made ofmaterials well-known in the pharmaceutical arts, for example, gelatin.However, in some embodiments, the delivery vehicle is integral with thepharmaceutical composition (i.e., the pharmaceutical composition is thedelivery vehicle). In such embodiments the pharmaceutical compositionscan be an insert, a gel, a cream, an ointment, a tablet, a suppository,or another preparation that is directly applied and absorbed vaginally.

In certain embodiments, the capsules do not contain one or more of thefollowing: a hydrophilic gel-forming bioadhesive agent, a lipophilicagent, a gelling agent for the lipophilic agent, and/or ahydrodispersible agent. In certain embodiments, the capsules do notcontain a hydrophilic gel-forming bioadhesive agent selected from:carboxyvinylic acid, hydroxypropylcellulose, carboxymethylcellulose,gelatin, xanthan gum, guar gum, aluminum silicate, and mixtures thereof.In certain embodiments, the capsules do not contain a lipophilic agentselected from: a liquid triglyceride, a solid triglyceride (with amelting point of about 35° C.), carnauba wax, cocoa butter, or mixturesthereof. In certain embodiments, the capsules do not contain ahydrophobic colloidal silica gelling agent. In certain embodiments, thecapsules do not contain a hydrodispersible agent selected from:polyoxyethylene glycol, polyoxyethylene glycol 7-glyceryl-cocoate, ormixtures thereof. In some such embodiments, the liquid composition canbe contained with a gelatin capsule as described herein. In some suchembodiments, the capsule can comprise gelatin and optionally one or morefurther components selected from the group consisting of gelatin,hydrolyzed gelatin, sorbitol-sorbitan solution, water, glycerin,titanium dioxide, FD&C Red #40, ethanol, ethyl acetate, propyleneglycol, polyvinyl acetate phthalate, isopropyl alcohol, polyethyleneglycol, and ammonium hydroxide.

In certain embodiments, the delivery vehicle can be designed for ease ofinsertion. In certain embodiments, the delivery vehicle can be sized sothat it can be comfortably inserted into the vagina. The deliveryvehicle can be prepared in a variety of geometries. For example, thedelivery vehicle can be shaped as a tear drop, a cone with frustoconicalends, a cylinder, a cylinder with larger “cap” portion, or in any othershape suitable for, and that eases insertion into, the vagina. Incertain embodiments, the delivery vehicle can be used in connection withan applicator. In other embodiments, the delivery vehicle can bedigitally inserted.

With reference to FIG. 2 , delivery vehicle 200 includes pharmaceuticalcomposition 202 and capsule 204. Width 208 represents the thickness ofcapsule 204, for example about 0.108 inches. The distance from one endof delivery vehicle 200 to another is represented by distance 206, forexample about 0.690 inches. The size of delivery vehicle 200 can also bedescribed by the arc swept by a radius of a given length. For example,arc 210, which is defined by the exterior of gelatin 204, is an arcswept by a radius of about 0.189 inches. Arc 212, which is defined bythe interior of capsule 204, is an arc swept by a radius of about 0.0938inches. Arc 214, which is defined by the exterior of gelatin 204opposite arc 210, is an arc swept by a radius of about 0.108 inches.Suitable capsules of other dimensions can be provided. In certainembodiments, capsule 204 has dimensions the same as or similar to theratios as provided above relative to each other. In some embodiments,the gelatin capsule can further comprise one or more components selectedfrom the group consisting of hydrolyzed gelatin, sorbitol-sorbitansolution, water, glycerin, titanium dioxide, FD&C Red #40, ethanol,ethyl acetate, propylene glycol, polyvinyl acetate phthalate, isopropylalcohol, polyethylene glycol, and ammonium hydroxide.

In certain embodiments, the delivery vehicle can be designed to remainin the vagina until the pharmaceutical composition is released. Incertain embodiments, the delivery vehicle can dissolve intravaginallyand can be absorbed into the vaginal tissue with the pharmaceuticalcomposition, which can minimize vaginal discharge. In some embodiments,the delivery vehicle can be made from constituents that are nontoxic,for example, gelatin.

EXAMPLES

The pharmaceutical compositions described herein are now furtherdetailed with reference to the following examples. These examples areprovided for the purpose of illustration only and the embodimentsdescribed herein should in no way be construed as being limited to theseexamples. Rather, the embodiments should be construed to encompass anyand all variations which become evident as a result of the teachingprovided herein.

Example 1: Solubilizing Agents

Compositions having the ingredients shown in Table 1 were prepared bycombining the ingredients using standard preparatory techniques.

TABLE 1 Estradiol Fill Formulas A B C Viscosity (cP) Composition/ 1418semi-solid 37.5 Component % w/w % w/w % w/w Estradiol 0.008% w/w  0.008% w/w   0.008% w/w   Triacetin 90% w/w Kolliphor HS 15 90% w/w(Solutol HS 15) Lauroglycol 90 90% w/w Tefose 63 10% w/w 10% w/wKolliphor RH40 10% w/w

Example 2: Pharmaceutical Composition

Estradiol was combined with one or more pharmaceutically acceptablesolubilizing agents. The estradiol used in the pharmaceuticalcomposition can be pharmaceutical grade, micronized estradiol, althoughother forms can also be used. The pharmaceutical composition includedestradiol in a dosage strength of from about 1 μg to about 50 μg. Thepharmaceutical composition can include 4 μg of estradiol. Thepharmaceutical composition can include 10 μg of estradiol. Thepharmaceutical composition can include 25 μg of estradiol.

The estradiol can be combined with pharmaceutically acceptablesolubilizing agents as described herein, and, optionally, otherexcipients, to form a pharmaceutical composition.

Pharmaceutical compositions comprising one or more solubilizing agentsas described herein that are liquid or semi-solid at room temperaturecan be tested using a Brookfield viscometer (Brookfield EngineeringLaboratories, Middleboro, Mass.) at room temperature.

Pharmaceutical compositions comprising one or more solubilizing agentsas described herein that are solid at room temperature can be assessedat 37° C. to determine their melting characteristics. The viscosity ofthe gels can be important during encapsulation of the formulation.

A dispersion assessment of the pharmaceutical compositions comprisingone or more solubilizing agents as described herein can be performed. Adispersion assessment can be performed by transferring 300 mg of eachvehicle system in 100 mL of 37° C. water, without agitation, andobserving for mixing characteristics. Generally speaking, it is believedthat formulations able to readily disperse in aqueous solution will havebetter dispersion characteristics upon administration.

Example 3: Delivery Vehicle

The pharmaceutical composition is delivered in a gelatin capsuledelivery vehicle. The gelatin capsule delivery vehicle includes, forexample, gelatin (e.g., Gelatin, NF (150 Bloom, Type B)), hydrolyzedcollagen (e.g., GELITA, GELITA AG, Eberbach, Germany), glycerin,sorbitol special, or other excipients in proportions that are well knownand understood by persons of ordinary skill in the art. Sorbitol specialcan be obtained commercially and can tend to act as a plasticizer andhumectant.

A variety of delivery vehicles, Gels A through F, were developed, asshown in Table 4.

TABLE 2 Gelatin Capsule Delivery Vehicles A B C D E F Ingredient % w/w %w/w % w/w % w/w % w/w % w/w Gelatin, NF 41.0 41.0 41.0 41.0 43.0 43.0(150 Bloom, Type B) Glycerin 99.7%, 6.0 6.0 6.0 6.0 18.0 18.0 USPSorbitol Special, 15.0 15.0 15.0 15.0 — — USP GELITA 3 — — — 3.0 —(hydrolyzed collagen) Citric acid — 0.1 0.5 1 — 0.1 Purified Water 35.037.9 37.5 37.0 36.0 38.9 TOTAL 100.0 100.0 100.0 100.0 100.0 100.0Dissolution gel 48 min 50 min 75 min 70 min — — strips, Avg. of (42, 45,(50, 51, (76, 75, (70, 71, 3 (500 mL, 58) 50) 74) 70) DH₂O, 50 rpm @ 37°C.) Dissolution gel 70 min — — — 78 min 82 min strips, Avg. of 3 (500mL, pH 4 buffer, 50 rpm @ 37° C.)

Each delivery vehicle A through F was prepared at a temperature rangefrom about 45° C. to about 85° C. Each molten delivery vehicle A throughF was cast into a film, dried, and cut into strips. The strips were cutinto uniform pieces weighing about 0.5 g, with about 0.5 mm thickness.Strips were placed into a USP Type 2 dissolution vessel in either wateror pH 4 buffer solution and the time for them to completely dissolve wasrecorded (see Table 4). Delivery vehicle A had the fastest dissolutionin both water and pH 4 buffer solution.

Example 4: Pharmaceutical Compositions with Delivery Vehicle

Various combinations of the pharmaceutical compositions comprising oneor more solubilizing agents described herein and delivery vehicles ofTable 4 can be prepared.

An aliquot of about 300 mg to about 310 mg of a pharmaceuticalcomposition comprising one or more solubilizing agents as describedherein can be prepared as described above. To encapsulate the vehiclesystem, each 300 mg to about 310 mg pharmaceutical composition aliquotcan be encapsulated in about 200 mg of the gelatin capsule deliveryvehicle. The aliquot size is arbitrary depending on the concentration ofthe estradiol and the desired gelatin capsule delivery vehicle size.Artisans will readily understand how to adjust the amount of estradiolin the pharmaceutical composition to accommodate a given size ofdelivery vehicle, when the delivery vehicle encapsulates thepharmaceutical composition.

Example 5: Process Using One or More Solubilizing Agents of the PresentDisclosure

FIG. 1 illustrates an embodiment of a method making a pharmaceuticalcomposition comprising estradiol solubilized in a solubilizing agentcomprising one or more solubilizing agents as described hereinencapsulated in a soft gelatin delivery vehicle 100. In operation 102,the solubilizing agent(s) is heated to 40° C.±5° C. Heating can beaccomplished through any suitable means. The heating can be performed inany suitable vessel, such as a stainless-steel vessel. Otherpharmaceutical compositions can be made using the same general method bysubstituting various excipients, including the solubilizing agent.

In operation 104, GELUCIRE is mixed with the first solubilizing agentcomprising to form the finished solubilizing agent. As used herein, anyof the second solubilizing agents described herein can be used inoperation 104 in place of GELUCIRE. Mixing is performed as would beknown to persons of ordinary skill in the art, for example by impeller,agitator, stirrer, or other like devices used to mix pharmaceuticalcompositions. Operation 104 can be performed under an inert orrelatively inert gas atmosphere, such as nitrogen gas. Mixing can beperformed in any vessels that are known to persons of ordinary skill inthe art, such as a stainless-steel vessel or a steel tank.

In operation 106 estradiol is mixed into the solubilizing agent. Inembodiments, the estradiol is in a micronized form when mixed into thesolubilizing agent. In other embodiments, the estradiol added is in anon-micronized form. Mixing can be facilitated by an impeller, agitator,stirrer, or other like devices used to mix pharmaceutical compositions.Operation 106 can be performed under an inert or relatively inert gasatmosphere, such as nitrogen gas.

In embodiments, however, the addition of estradiol can be performedprior to operation 104. In that regard, operations 104 and 106 areinterchangeable with respect to timing or can be performedcontemporaneously with each other.

In operation 110, the gelatin delivery vehicle is prepared. Any of thegelatin delivery vehicles described herein can be used in operation 110.In embodiments, gelatin, hydrolyzed collagen, glycerin, and otherexcipients are combined at a temperature range from about 45° C. toabout 85° C. and prepared as a film. Mixing can occur in a steel tank orother container used for preparing gelatin delivery vehicles. Mixing canbe facilitated by an impellor, agitator, stirrer, or other devices usedto combine the contents of gelatin delivery vehicles. Operation 110 canbe performed under an inert or relatively inert gas atmosphere, such asnitrogen gas. In embodiments, the gelatin delivery vehicle mixture isdegassed prior to being used to encapsulate the pharmaceuticalcomposition.

In operation 112, the gelatin delivery vehicle encapsulates thepharmaceutical composition, according to protocols well-known to personsof ordinary skill in the art. In operation 112, a soft gelatin capsuledelivery vehicle is prepared by combining the pharmaceutical compositionmade in operation 106 with the gelatin delivery vehicle made inoperation 110. The gelatin can be wrapped around the material, partiallyor fully encapsulating it or the gelatin can also be injected orotherwise filled with the pharmaceutical composition made in operation106.

In embodiments, operation 112 is completed in a suitable die to providea desired shape. Vaginal soft gel capsules can be prepared in a varietyof geometries. For example, vaginal soft gel capsules can be shaped as atear drop, a cone with frustoconical end, a cylinder, a cylinder withlarger “cap” portion as illustrated in FIG. 2 , or other shapes suitablefor insertion into the vagina. The resulting pharmaceutical compositionencapsulated in the soft gelatin delivery vehicle can be inserteddigitally or with an applicator.

The breadth and scope of the present invention should not be limited byany of the above-described exemplary embodiments but should be definedonly in accordance with the following claims and their equivalents.

All patents, patent applications, and other references noted orreferenced in this application are hereby incorporated by reference intheir entirety.

1. A vaginal insert comprising a therapeutically effective amount of estradiol; and a solubilizing agent, wherein the solubilizing agent comprises at least one C₂-C₅ fatty acid or a glycol ester, monoglyceride, diglyceride, or triglyceride thereof.
 2. The vaginal insert of claim 1, wherein the therapeutically effective amount of estradiol is from about 1 microgram to about 25 micrograms. 3.-6. (canceled)
 7. The vaginal insert of claim 1, further comprising a second solubilizing agent selected from the group consisting of a mixture of PEG-6 stearate, ethylene glycol palmitostearate, and PEG-32 stearate; a mixture of hard fat, glyceryl ricinoleate, ceteth-20, and steareth-20; polyoxyl 40 hydrogenated castor oil USP; hard fat polyoxyl 20 cetostearyl ether; and combinations thereof.
 8. The vaginal insert of claim 7, wherein the second solubilizing agent is a mixture of PEG-6 stearate, ethylene glycol palmitostearate, and PEG-32 stearate.
 9. The vaginal insert of claim 1, wherein the viscosity of the composition is about 5 cP to about 3000 cP at room temperature.
 10. (canceled)
 11. The vaginal insert of claim 1, wherein the insert further comprises a capsule encapsulating the therapeutically effective amount of estradiol and the solubilizing agent.
 12. (canceled)
 13. The vaginal insert of claim 1, wherein the solubilizing agent comprises at least one C₃ fatty acid or a glycol mono- or di-ester thereof, a monoglyceride, diglyceride, or triglyceride thereof, or a combination of any of the foregoing.
 14. A vaginal insert comprising: (a) a therapeutically effective amount of estradiol; (b) a first solubilizing agent comprising one or more polyethylene glycol mono- or di-esters of a hydroxy C₁₆-C₂₆ fatty acid; and (c) a second solubilizing agent comprising a mixture of PEG-6 stearate, ethylene glycol palmitostearate, and PEG-32 stearate.
 15. The vaginal insert of claim 14, wherein the therapeutically effective amount of estradiol is from about 1 microgram to about 25 micrograms. 16.-20. (canceled)
 21. The vaginal insert of claim 14, wherein the insert further comprises a capsule encapsulating the estradiol, the first solubilizing agent, and the second solubilizing agent.
 22. (canceled)
 23. A vaginal insert comprising: (d) a therapeutically effective amount of estradiol; (e) a first solubilizing agent comprising propylene glycol monolaurate; and (f) a second solubilizing agent comprising polyoxyl 40 hydrogenated castor oil USP.
 24. The vaginal insert of claim 23, wherein the therapeutically effective amount of estradiol is from about 1 microgram to about 25 micrograms. 25.-28. (canceled)
 29. The vaginal insert of claim 23, wherein the viscosity of the composition is about 5 cP to about 3000 cP at room temperature.
 30. (canceled)
 31. The vaginal insert of claim 23, wherein the insert further comprises a capsule encapsulating the estradiol, the first solubilizing agent, and the second solubilizing agent.
 32. (canceled)
 33. A method of treating an estrogen-deficient state, comprising administering to a female in need thereof, the vaginal insert of claim
 1. 34. (canceled)
 35. The method of claim 33, wherein the estrogen-deficient state is selected from the group consisting of vulvovaginal atrophy, dysuria, dyspareunia, estrogen-deficient urinary state, and vaginal bleeding associated with sexual activity.
 36. (canceled)
 37. A method of treating an estrogen-deficient state, comprising administering to a female in need thereof, the vaginal insert of claim
 14. 38. (canceled)
 39. The method of claim 37, wherein the estrogen-deficient state is selected from the group consisting of vulvovaginal atrophy, dysuria, dyspareunia, estrogen-deficient urinary state, and vaginal bleeding associated with sexual activity.
 40. (canceled)
 41. A method of treating an estrogen-deficient state, comprising administering to a female in need thereof, the vaginal insert of claim
 23. 42. (canceled)
 43. The method of claim 41, wherein the estrogen-deficient state is selected from the group consisting of vulvovaginal atrophy, dysuria, dyspareunia, estrogen-deficient urinary state, and vaginal bleeding associated with sexual activity.
 44. (canceled) 